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<p>Garlic and onion, key <i>Allium</i> crops, are cultivated globally for culinary and medicinal purposes. Onion yellow dwarf virus (OYDV) is a major pathogen affecting these crops, causing leaf crinkling, yellow striping, dwarfing, and reduced yield. To facilitate efficient detection, a rapid, reliable, and cost-effective crude leaf extract-based reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed. This method eliminates the need for labour-intensive RNA isolation, cDNA synthesis, and expensive thermal cycler used in PCR/RT-PCR based assay. The optimized RT-RPA assay showed comparable sensitivity to RT-PCR, detecting OYDV up to 0.1 pg/μl, or 10<sup>-6</sup> using RNA or crude leaf extract. It exhibited high specificity, with no cross-reactions with other <i>Allium</i> viruses. This study presents the first crude leaf extract-based RT-RPA test for OYDV detection, offering a valuable tool for routine indexing, biosecurity, and resistance screening.</p>

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A crude leaf extract-based RT-RPA assay for rapid detection of onion yellow dwarf virus in Allium crops

  • 期刊论文

  • 发表日期：

    2025年12月1日

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  • Rakesh Kumar,
  • Nitika Gupta,
  • Susheel Kumar Sharma,
  • Nishant Srivastava,
  • Pooja Bhardwaj,
  • Sonia Kapoor,
  • Rajendra Prasad Pant,
  • Virendra Kumar Baranwal

摘要

Garlic and onion, key Allium crops, are cultivated globally for culinary and medicinal purposes. Onion yellow dwarf virus (OYDV) is a major pathogen affecting these crops, causing leaf crinkling, yellow striping, dwarfing, and reduced yield. To facilitate efficient detection, a rapid, reliable, and cost-effective crude leaf extract-based reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed. This method eliminates the need for labour-intensive RNA isolation, cDNA synthesis, and expensive thermal cycler used in PCR/RT-PCR based assay. The optimized RT-RPA assay showed comparable sensitivity to RT-PCR, detecting OYDV up to 0.1 pg/μl, or 10-6 using RNA or crude leaf extract. It exhibited high specificity, with no cross-reactions with other Allium viruses. This study presents the first crude leaf extract-based RT-RPA test for OYDV detection, offering a valuable tool for routine indexing, biosecurity, and resistance screening.